Figure 3: A dynamic actin meshwork forms around the nucleus and deform this organelle during its passage through narrow constrictions. | Nature Communications

Figure 3: A dynamic actin meshwork forms around the nucleus and deform this organelle during its passage through narrow constrictions.

From: Perinuclear Arp2/3-driven actin polymerization enables nuclear deformation to facilitate cell migration through complex environments

Figure 3

(a) Sequential images of a representative LifeAct-GFP (green) expressing DC stained with Hoechst (red) passing a 2-μm-wide constriction. Scale bar, 20 μm. (b) Normalized mean actin intensity around the nucleus (green) and nuclear circularity (red) of cells crossing 2-μm-wide constrictions as a function of the nuclear centre of mass position relative to the centre of the constriction. (c) Normalized mean actin intensity around the nucleus in 1.5 μm (dark green) and 5 μm (light green) wide constrictions as a function of the nuclear centre of mass position relative to the centre of the constriction. (d) Immunostaining of a DC migrating in a collagen gel; yellow arrows show actin accumulation at the site of nuclear deformation. Scale bar, 10 μm (left); 5 μm (right). (e) RFP-LifeAct (green)- and H2B-CFP (red)-expressing myeloid cells migrating in mekada fish during in vivo wound healing. Yellow arrows indicate actin accumulation around the deformed nucleus (orange arrows, representative of 53 of the 66 observed cells). Time frames in hours: minutes: seconds. Scale bar, 15 μm. (f) Normalized mean actin intensity at the nuclear region in myeloid cells migrating in mekada fish as a function of nuclear circularity. Error bars are s.e.m. (g) Mean actin intensity as a function of DNA thickness at a position X in 1.5-μm-wide constrictions. Red line indicates the linear regression of the data for DNA tickness smaller than 1.5 μm (the constriction width). Error bars are s.d. (h) Mean actin intensity as a function of mean DNA intensity at a position X in 1.5-μm-wide constrictions. Red line indicates the linear regression of the data for DNA mean intensity smaller than 2,500 AU μm−2. Error bars are s.e.m. (i) Representative spinning disc images of LifeAct-GFP-expressing (actin, green) DC stained with Hoechst (DNA, red) in 2-μm-wide constrictions. Scale bar, 10 μm. White arrow indicates the position of the line scan. Scale bar, 2 μm. L, constriction length; n, number of cells; W, constriction width. N≥2.

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